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A comprehensive investigation of the medicinal efficacy of antimicrobial fusion peptides expressed in probiotic bacteria for the treatment of pan drug-resistant (PDR) infections

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Abstract

The present work aimed to examine the intracellular antibacterial efficacy of Recombinant Lactobacillus acidophilus/antimicrobial peptides (AMPs) Melittin and Alyteserin-1a, specifically targeting Gram-negative bacteria. The first assessment was to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Recombinant L. acidophilus/AMPs versus Gram-negative and Gram-positive bacteria. In addition, the researchers examined the in vitro viability and safety of AMPs generated by L. acidophilus. The experiments included exposing the AMPs to elevated temperatures, proteases, cationic salts at physiological levels, and specific pH settings. The safety aspect was evaluated using hemolytic analysis utilizing sheep erythrocytes; cytotoxicity assays employing cell lines, and experiments on beneficial gut lactobacilli. An experiment was done using a time-kill method to assess the intracellular antibacterial efficacy of Recombinant L. acidophilus/AMPs compared to pathogenic varieties in HEp-2 cells. Previous investigations have shown that the MBC levels of recombinant L. acidophilus/AMPs were consistently two to four times higher than the equivalent MIC values when evaluated versus Gram-negative bacteria. Furthermore, the stability of the Recombinant L. acidophilus/AMPs showed variability when exposed to elevated temperatures (70 and 90 ℃), treated with protease enzymes (proteinase K, lysozyme), exposed to higher concentrations of physiological salts (150 mM NaCl and 2 mM MgCl2), and varying pH levels (ranging from 4.0 to 9.0). The recombinant L. acidophilus/AMPs are non-hemolytic towards sheep erythrocytes, exhibit little cytotoxicity in RAW 264.7 and HEp-2 cells, and are considered safe when compared to beneficial gut lactobacilli. The research examined the intracellular bacteriostatic effects of recombinant L. acidophilus/AMPs on Gram-negative bacteria inside HEp-2 cells. Nevertheless, no notable bactericidal impact was seen on Gram-positive bacteria (P > 0.05). The research shows that recombinant L. acidophilus/AMPs, namely (L. acidophilus/melittin/Alyteserin-1a) as the focus of the investigation, effectively eliminate Gram-negative bacteria. Therefore, more investigation is necessary to elaborate on these discoveries.

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Data availability

The data sets generated and/or analyzed during the current study are available in the https://www.uniprot.org/uniprotkb/Q8LW54/entry.

Abbreviations

MIC:

Minimum inhibitory concentration

MBC:

Minimum bactericidal concentration

MDR:

Multidrug resistance

XDR:

Extensive drug resistance

ESBLs:

Extended-spectrum beta-lactamases

CRE:

Carbapenem-resistant Enterobacteriaceae

MRSA:

Methicillin-resistant S. aureus

PDR:

Pan drug resistance

CAMA:

Cecropin A (1–7)-Melittin

LAB:

Lactic acid bacteria

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Acknowledgements

The authors would like to thank the staff members of the Biotechnology Research Center of the Islamic Azad University of Shahrekord Branch in Iran for their help and support. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

    Maryam Safarpour-Dehkordi, Omid Chabok, Mohsen Asgari & Reyhaneh Khademi

  2. Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

    Abbas Doosti

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  1. Maryam Safarpour-Dehkordi
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  2. Omid Chabok
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Contributions

MSD conducted research and drafted the manuscript, AD conceived and designed research and made manuscript revision, OC, MA and RK designed research and analyzed data, AD provided funding support. All authors read and approved the final manuscript.

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Correspondence to Abbas Doosti.

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Safarpour-Dehkordi, M., Chabok, O., Asgari, M. et al. A comprehensive investigation of the medicinal efficacy of antimicrobial fusion peptides expressed in probiotic bacteria for the treatment of pan drug-resistant (PDR) infections. Arch Microbiol 206, 93 (2024). https://doi.org/10.1007/s00203-023-03823-2

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